Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Mannoproteins released during the fermentation of Saccharomyces cerevisiae in a chemically defined synthetic grape juice medium were isolated by ethanol precipitation. The proteins from the medium were fractionated by ion exchange chromatography. Fractions containing mannoproteins were identified by their UV absorption spectra and by the presence of polymeric mannose. A mannoprotein designated HPF2 was purified from one of the fractions by gel permeation chromatography, and had a high capacity to reduce heat-induced protein haze formation in white wine. After electrophoretic separation and transfer to nitrocellulose, HPF2 stained positively with an antibody (anti-HPFl) which had been raised against a previously isolated mannoprotein with known haze protective activity designated HPFL. Analysis of the carbohydrate portion of HPF2 using methylation analysis confirmed the presence of (1 -> 2), (1 -> 3) and (1 -> 2,1 -> 6) mannosyl residues and showed that it contained N-linked and possibly O-linked carbohydrate. Digestion of the mannoprotein with PNGase F resulted in a reduction in molecular weight, as measured by SDS-PAOE and confirmed the presence of N-linked carbohydrate. N-linked deglycosylation decreased the ability of HPF2 to protect wine from heatinduced protein haze. Protein sequence analysis of the peptides derived from the HPF2 mannoprotein obtained via digestion with endoproteinase Lys C led to the identification of the HPF2 structural gene in Saccharomyces cerevisiae followed by searching the Yeast Proteome Database for related sequences. Analysis of the deduced amino acid sequence of HPF2 from its structural gene indicated that HPF2 possessed a secretion signal at the NH₂-terminus, a putative OPI anchor site at the COOH-terminus and also contained serine and threonine rich regions at both the NH₂-terminus and COOHterminus. These regions may contain O-linked carbohydrate. There were also 15 potential glycosylation sites, five of which were classified from the peptide mapping data as likely glycosylation sites. These characteristics, combined with the results from the carbohydrate analysis, indicate that HPF2 was a mannoprotein derived from the yeast cell walls. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=678380 / Thesis (Ph.D.) -- University of Adelaide, Dept. of Plant Science, 2000

Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Mannoproteins released during the fermentation of Saccharomyces cerevisiae in a chemically defined synthetic grape juice medium were isolated by ethanol precipitation. The proteins from the medium were fractionated by ion exchange chromatography. Fractions containing mannoproteins were identified by their UV absorption spectra and by the presence of polymeric mannose. A mannoprotein designated HPF2 was purified from one of the fractions by gel permeation chromatography, and had a high capacity to reduce heat-induced protein haze formation in white wine. After electrophoretic separation and transfer to nitrocellulose, HPF2 stained positively with an antibody (anti-HPFl) which had been raised against a previously isolated mannoprotein with known haze protective activity designated HPFL. Analysis of the carbohydrate portion of HPF2 using methylation analysis confirmed the presence of (1 -> 2), (1 -> 3) and (1 -> 2,1 -> 6) mannosyl residues and showed that it contained N-linked and possibly O-linked carbohydrate. Digestion of the mannoprotein with PNGase F resulted in a reduction in molecular weight, as measured by SDS-PAOE and confirmed the presence of N-linked carbohydrate. N-linked deglycosylation decreased the ability of HPF2 to protect wine from heatinduced protein haze. Protein sequence analysis of the peptides derived from the HPF2 mannoprotein obtained via digestion with endoproteinase Lys C led to the identification of the HPF2 structural gene in Saccharomyces cerevisiae followed by searching the Yeast Proteome Database for related sequences. Analysis of the deduced amino acid sequence of HPF2 from its structural gene indicated that HPF2 possessed a secretion signal at the NH₂-terminus, a putative OPI anchor site at the COOH-terminus and also contained serine and threonine rich regions at both the NH₂-terminus and COOHterminus. These regions may contain O-linked carbohydrate. There were also 15 potential glycosylation sites, five of which were classified from the peptide mapping data as likely glycosylation sites. These characteristics, combined with the results from the carbohydrate analysis, indicate that HPF2 was a mannoprotein derived from the yeast cell walls. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=678380 / Thesis (Ph.D.) -- University of Adelaide, Dept. of Plant Science, 2000

Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Mannoproteins released during the fermentation of Saccharomyces cerevisiae in a chemically defined synthetic grape juice medium were isolated by ethanol precipitation. The proteins from the medium were fractionated by ion exchange chromatography. Fractions containing mannoproteins were identified by their UV absorption spectra and by the presence of polymeric mannose. A mannoprotein designated HPF2 was purified from one of the fractions by gel permeation chromatography, and had a high capacity to reduce heat-induced protein haze formation in white wine. After electrophoretic separation and transfer to nitrocellulose, HPF2 stained positively with an antibody (anti-HPFl) which had been raised against a previously isolated mannoprotein with known haze protective activity designated HPFL. Analysis of the carbohydrate portion of HPF2 using methylation analysis confirmed the presence of (1 -> 2), (1 -> 3) and (1 -> 2,1 -> 6) mannosyl residues and showed that it contained N-linked and possibly O-linked carbohydrate. Digestion of the mannoprotein with PNGase F resulted in a reduction in molecular weight, as measured by SDS-PAOE and confirmed the presence of N-linked carbohydrate. N-linked deglycosylation decreased the ability of HPF2 to protect wine from heatinduced protein haze. Protein sequence analysis of the peptides derived from the HPF2 mannoprotein obtained via digestion with endoproteinase Lys C led to the identification of the HPF2 structural gene in Saccharomyces cerevisiae followed by searching the Yeast Proteome Database for related sequences. Analysis of the deduced amino acid sequence of HPF2 from its structural gene indicated that HPF2 possessed a secretion signal at the NH₂-terminus, a putative OPI anchor site at the COOH-terminus and also contained serine and threonine rich regions at both the NH₂-terminus and COOHterminus. These regions may contain O-linked carbohydrate. There were also 15 potential glycosylation sites, five of which were classified from the peptide mapping data as likely glycosylation sites. These characteristics, combined with the results from the carbohydrate analysis, indicate that HPF2 was a mannoprotein derived from the yeast cell walls. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=678380 / Thesis (Ph.D.) -- University of Adelaide, Dept. of Plant Science, 2000

Abstract The competition for liquor companies in Taiwan is getting fierce due to limited market size and tardy economic growth. The situation is more obvious after Taiwan joined WTO and many famous foreign liquor brands entered this market. The impact is especially strong for Chinese Spirits because many liquor consumers take a shot at these new entrants or even change their habits. Kinmen Kaoliang Liquor Inc. now faces domestic competitors such as Yusan Kaoliang of Taiwan Tabacco and Liquor Coporation and other brands as well as the growing portion of consumers shifting to bear or wine. In order to operate in China and seek further revenue growth, Kinmen Kaoliang Liquor Inc. established a child company in Xiamen in 2004. However, it is shown that there¡¦s much room for further effort if the company wants to perform well in China. This research adopts exploratory research method and collects relevant data from various sources. Through sorting, estimating, evaluating, and reasoning of the data, this research provides workable strategies for Kinmen Kaoliang Liquor Inc. for further development. Keyword¡Gwhite wine, Kinmen Kaoliang Liquor, Marketing Strategy, target market

No description available.

Effectiveness of a capture and return system for the partial retention of fermentation volatiles, as a means of improving white wine quality, was evaluated. Twenty-three aroma-active volatiles including ethyl esters, acetate esters, fusel alcohols, and fatty acids, were quantified using head-space solid phase microextraction with GC/MS. Volatile analysis of fermentations maintained at 15ºC demonstrated a trend of increased concentrations of ethanol, esters and ethyl esters of fatty acids and decreased concentrations of fusel alcohol acetates, fatty acids and higher alcohols in treatment wines. When fermentation temperature was maintained at 15ºC there was increased concentration and retention of fusel alcohols, fatty acids and higher alcohols compared to 15ºC. Sensory analysis of wines fermented at 15°C, using triangle difference testing, indicated variable differences in aroma among treatments. / Master of Science

Les réactions d'oxydation impliquant les composés phénoliques semblent induire des modifications non négligeables du profil chimique et sensoriel des vins. Les travaux concernent l’étude des mécanismes réactionnels impliquant certains thiols volatils, contributeurs de l’arôme distinctif et de la complexité des vins des différents cépages avec les composés phénoliques oxydés des vins blancs, principalement les flavan-3-ols. En solution modèle de composition proche du vin, une réactivité différente des thiols volatils selon leur nature chimique vis à vis des formes oxydées des flavan-3-ols a été établie. La synthèse et la caractérisation des adduits par RMN entre les principaux composés phénoliques des moûts et des vins blancs et le 3-sulfanylhexanol, présentant des nuances d’agrumes, a ensuite été réalisée en conditions d’oxydation chimique et enzymatique. La suivi cinétique de la formation des adduits par CLHP-ESI-SM a permis de mettre en évidence une réactivité du thiol spécifique vis à vis d’un substrat polyphénolique, d’établir le rôle catalytique des métaux (Fe2+) et la capacité antioxydante du dioxyde de soufre vis à vis de ces mécanismes réactionnels. La compréhension de mécanismes fondamentaux de la réactivité de la (+)-catéchine et de la (-)-épicatéchine en conditions œnologiques avec les thiols volatils nous a permis de décliner les travaux à l’étude de l’influence de la présence des flavan-3-ols au cours de la vinification et de l’élevage des vins sur ces composants de l’arôme des vins blancs. / Oxidation reactions involving phenolics might change wines chemical and sensory profile. The present work concern the study of reactional mechanisms implying certain volatile thiols, responsible of distinctiveness and complexity of various wines, with white wines oxidized phenolic compounds, mainly flavan-3-ols. In a model wine solution, a different volatile thiol reactivity pattern according to their chemical nature with respect to oxidized flavan-3-ols forms was established. The adducts synthesis and characterization by NMR between the principal white musts and wines phenolic compounds and the 3-sulfanylhexanol, presenting citrus fruits nuances, were carried out under chemical and enzymatic oxidation conditions. Their formation monitoring by HPLC-ESI-MS highlighted a specific reactivity of thiol with polyphenolic substrate and established the catalytic role of metals (Fe2+) as well as, the antioxidant effect of sulphur dioxide into these mechanisms. The comprehension of fundamental mechanisms for the reactivity of (+)-catechin and of (-)-epicatechin with volatile thiols in oenological conditions enabled us to elucidate the influence of flavan-3-ols into white wines aroma compounds during wine making and ageing.

Las proteínas térmicamente inestables que están presentes en uvas, jugos de uva y vinos podrían llegar a ser insolubles y precipitar causando la formación de turbidez o precipitados indeseables en vinos blancos después del embotellado durante el almacenamiento.La turbidez proteica en vinos blancos es evitada tradicionalmente, mediante la adición de bentonita, aunque esta técnica presenta algunas desventajas tales como efectos negativos sobre las propiedades sensoriales del vino debido principalmente a la remoción de componentes aromáticos o gustativos y por la merma de vino, debido al gran aumento de volumen y poder de sedimentación de la bentonita. Además, el recurso humano, los tiempos de proceso y la descarga de residuos al ambiente sigue siendo una inmensa preocupación, debido a los significativos costos asociados a la salud y seguridad laboral, y así como también las responsabilidades y obligaciones legales de la industria en material de impacto ambiental. Es estimado que los costos del uso de la bentonita en la industria del vino a nivel mundial son del orden de los 300-500 millones de dólares por año. Por lo tanto es necesario el desarrollo de nuevas tecnologías alternativas a la bentonita que sean económicamente viables y que mantengan la calidad del vino, así como también la generación de un menor impacto ambiental. No obstante, nuevas técnicas exitosas a nivel industrial aun no han sido desarrolladas, por que afectan la calidad del vino o porque su aplicación no es viable económicamente bajo normales condiciones de operación de producción de vino. Por lo tanto, es muy atractivo investigar la viabilidad de nuevas prácticas que tengan un menor impacto sobre el ambiente y sean económicamente viables.Por esa razón la principal motivación de nuestra investigación ha sido estudiar la viabilidad industrial de una tecnología alternativa al uso de la bentonita, la cual permita remover proteínas inestables de los vinos blancos usando zirconia como material adsorbente. Además, nosotros nos hemos concentrado en el desarrollo de un proceso continuo que permita conseguir vinos estables proteicamente sin afectar sus propiedades fisicoquímicas y sensoriales, probando diferentes técnicas de regeneración del material adsorbente. Primeramente nosotros hemos estudiado la estructura, morfología y propiedades superficiales de la zirconia, así como también su capacidad de adsorción para remover proteínas inestables de vinos blancos, aplicando tratamientos regenerativos químicos y térmicos. Después hemos comparado las propiedades fisicoquímicas y sensoriales de un vino blanco estabilizado proteicamente mediante zirconia y bentonita.Además, nosotros hemos desarrollado un proceso híbrido integrando un proceso de adsorción en columna y un proceso de microfiltración tangencial de vino, para conocer los efectos de este nuevo proceso sobre el ensuciamiento de la membrana y la estabilidad proteica del vino.Por otro lado, nosotros también hemos aplicado un nuevo proceso de estabilización proteica de vino base para cava, comparando los resultados con el método tradicional usando bentonita como agente estabilizante y observando los efectos de ambos tratamientos sobre la calidad de la espuma y las fracciones proteicas del vino. Finalmente, hemos aplicado el nuevo proceso de estabilización proteica a escala industrial, empacando zirconia sobre una columna fija y realizando el proceso mediante sistema continuo y discontinuo.Los resultados demuestran que la zirconia puede ser regenerada química o térmicamente, que sus propiedades físicas, morfológicas y químicas no son alteradas y que incluso su capacidad de adsorción proteica podría ser aumentada probablemente producto de la adsorción de algunos componentes o centros activos derivados de las proteínas del vino. A través del proceso híbrido ha sido posible conseguir vinos estables proteicamente y aumentar la densidad de flujo del permeado durante la microfiltración del vino. De hecho, hemos observado que la reducción de proteínas mediante la adsorción en columna usando zirconia también ocurre durante la microfiltración tangencial, por lo tanto ambos procesos pueden actuar conjuntamente en la reducción y estabilización proteica.Por otro lado, comparando las propiedades fisicoquímicas y sensoriales de vinos blancos estabilizados proteicamente mediante zirconia y bentonita, podemos afirmar que los mejores resultados son conseguidos usando zirconia como agente estabilizante.La estabilización proteica de vinos blancos en continuo también puede ser útil para estabilizar vinos base para cava, ya que comparando la calidad de la espuma de un vino base para cava tratado con zirconia y bentonita, los resultados demuestran que la calidad de la espuma de aquellos vinos bases es mejor usando zirconia, ya que la adicción de bentonita produce considerables efectos negativos sobe la calidad de la espuma.Finalmente, los resultados obtenidos a escala industrial, muestran que es viable la estabilización proteica de vinos blancos usando zirconia como material adsorbente ya sea mediante un sistema continuo o discontinuo sin afectar la calidad del vino inicial. / Heat-unstable soluble proteins in grapes, grape juices and wines may become insoluble and precipitate causing the formation of undesirable hazes or deposits in white wines after bottling and during storage. Proteins are commonly prevented from forming hazes with bentonite, though this technique does have drawbacks: for example, the sensory properties of the wine are affected adversely because flavour compounds are removed and wine volume is lost as lees because of the swell and settling of the bentonite. In addition, the handling and disposal of spent bentonite continues to be a concern, because it involves high labour input and the associated costs, occupational health and safety issues, and the wine industry's environmental responsibilities and legislative requirements. It is estimated that the cost of bentonite fining to the wine industry worldwide is in the order of US$300-500 m per annum. Alternative fining technologies to bentonite, which are economically viable and maintain wine quality, are currently being sought. However, no successful techniques have been developed to date: all the attempts so far have either affected the quality of the wine or not been economically viable under standard winemaking conditions. Therefore, research on the implementation of new practices that have a less negative impact on the environment and are economically viable is particularly challenging.For this reason the aim of this thesis was to study the industrial viability of an alternative technology to bentonite fining which enables unstable proteins to be removed from white wines using zirconia as the adsorbent material. We also attempted to develop a continuous process, which stabilizes wine protein without having any negative effects on the physicochemical and sensory properties of the wine. Likewise we tried to make the process have a lower environmental impact by testing various regenerative treatments of the adsorbent material.First we studied the structure, morphology and surface properties of the zirconia and its capacity to remove unstable proteins from white wine versus thermal and chemical regeneration treatments. Subsequently we compared the physicochemical and sensory properties of a white wine fined by zirconia and bentonite. To further our understanding of the effect on membrane fouling and wine protein stability, we developed a hybrid process consisting of in-column adsorption with crossflow microfiltration. We also applied this new method to stabilize the base sparkling wine and compared the results with the conventional method of using bentonite as the fining agent to see the effects of the treatments on the foam quality and protein fractions. Finally, we applied the new method on an industrial scale by packing zirconia into a fixed bed column and by using the batch and continuous systems.The results show that the zirconia can be regenerated by thermal and chemical treatments, and that its physical, morphological and chemical properties are not altered. In fact its protein adsorption capacity can increase probably because some compounds or active centres derived from wine proteins are absorbed.The hybrid process was used to increase the permeate flux during crossflow microfiltration and stabilize wine proteins. We observed that proteins were reduced when the zirconia column adsorption was used during the crossflow microfiltration. Therefore both processes may act together.By comparing the physicochemical and sensory analyses of white wine proteins stabilized by zirconia and bentonite, we found that results were best when zirconia was used.The continuous protein stabilization of white wines by zirconia may also be useful for stabilizing proteins in base sparkling wines. Treating base sparkling wines with zirconia definitely gives better foam quality than with bentonite. Finally the results obtained in our industrial scale experiment showed that white wine continuous protein stabilization with zirconia as the adsorbent material is not only viable in both the continuous and batch systems, it also leaves the quality of the wine unchanged.

Heat induced protein haze is a common problem in white wine. Grape derived pathogenesis related proteins slowly denature and aggregate during wine storage and this gives rise to light dispersing haze. Protein haze formation is currently prevented by removing proteins using bentonite, an aluminium silicate clay, but this method has drawbacks. A potential alternative or complementary method is the use of haze protective factors ( HPF ), specific mannoproteins from Saccharomyces cerevisiae that visually reduce protein haze. Hpf1p was originally isolated from Muscat Gordo Blanco wine and Hpf2p from a synthetic grape juice ferment. Based on partial amino acid sequences, putative structural genes, HPF1 and HPF2, for these proteins were identified. HPF1 has a homologue, HPF1 ', ( 71 % similarity ) in S. cerevisiae. Sequence analysis suggests that Hpf1p, Hpf1 ' p and Hpf2p are localised to the cell wall or plasma membrane. This study aimed to determine the biological function of the HPF genes in S. cerevisiae. HPF overexpression and deletion strains were constructed and analysed for cell wall related phenotypes. Under a number of conditions, including cold temperature and ethanol stress, the hpf1 Δ hpf1 ' Δ strain was more tolerant than the wild type strain. However, mating efficiency of the hpf1 Δ hpf1 ' Δ strain was significantly less than the wild type strain and this was found to be correlated with the persistence of a septum between the mating partners. The decreased mating efficiency was also mating type specific, only occurring in MAT α cells. This study also aimed to establish conclusively that the HPF genes do indeed encode proteins with haze protective properties. Haze protective activity of the material from ferment supernatants was assessed. Material from the HPF deletion strains exhibited significantly less haze protective activity than the wild type. Moreover, material derived from HPF1 and HPF1 ' overexpressors was more active than material from the wild type. A 6xHis - tagged Hpf2p was expressed and purified using immobilised metal affinity chromatography. This Hpf2p had significant haze protective activity. Modification of N - glycans of 6xHis - Hpf2p by Endoglycosidase H decreased its haze protective activity.visually reduce protein haze. Hpf1p was originally isolated from Muscat Gordo Blanco wine and Hpf2p from a synthetic grape juice ferment. Based on partial amino acid sequences, putative structural genes, HPF1 and HPF2, for these proteins were identified. HPF1 has a homologue, HPF1 ', ( 71 % similarity ) in S. cerevisiae. Sequence analysis suggests that Hpf1p, Hpf1 ' p and Hpf2p are localised to the cell wall or plasma membrane. This study aimed to determine the biological function of the HPF genes in S. cerevisiae. HPF overexpression and deletion strains were constructed and analysed for cell wall related phenotypes. Under a number of conditions, including cold temperature and ethanol stress, the hpf1 Δ hpf1 ' Δ strain was more tolerant than the wild type strain. However, mating efficiency of the hpf1 Δ hpf1 ' Δ strain was significantly less than the wild type strain and this was found to be correlated with the persistence of a septum between the mating partners. The decreased mating efficiency was also mating type specific, only occurring in MAT α cells. This study also aimed to establish conclusively that the HPF genes do indeed encode proteins with haze protective properties. Haze protective activity of the material from ferment supernatants was assessed. Material from the HPF deletion strains exhibited significantly less haze protective activity than the wild type. Moreover, material derived from HPF1 and HPF1 ' overexpressors was more active than material from the wild type. A 6xHis - tagged Hpf2p was expressed and purified using immobilised metal affinity chromatography. This Hpf2p had significant haze protective activity. Modification of N - glycans of 6xHis - Hpf2p by Endoglycosidase H decreased its haze protective activity. / Thesis (Ph.D.)--School of Agriculture and Wine, 2003.

The present thesis work is dedicated to the implementation of novel electrochemical methods for the assay of drug compounds such as paracetamol and ascorbic acid and biologically relevant biothiols such as cysteine and glutathione. Particular attention has been paid to the development and application of amperometric detectors allowing for a readily surface renewing and for highly selective and sensitive assays.<p>Coupling of a screen printed electrochemical detector with a monolithic chromatographic column was performed for high-throughput analysis along with selectivity, sensitivity and good precision. The modification of a glassy carbon electrode with a perm-selective membrane was investigated in order to provide an electrochemical sensor for on-site analysis. Acetaminophen was investigated as a model drug compound because of its extensive use in drug “pain killer” medication.<p>Along with the aims of this thesis, a silver polycrystalline electrode was investigated as a sensitive and class selective electrode for the assay of small thiol-based molecules. The silver electrode was implemented for the first time as an amperometric detector coupled to liquid chromatography for the assay of thiols. Application to the study of thiol species present in white wines was realized in order to illustrate the potential of the silver working electrode as amperometric detector. / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished